Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Flow Cytometry, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockout Validated, Western Blot

Cited:

Flow Cytometry, IF/IHC, Immunoblotting, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Immunoprecipitation, Knockout Validated, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

View all Pyruvate Dehydrogenase E1-alpha subunit Antibodies »

Product Specifications

Immunogen

A synthetic peptide surrounding the phosphorylated serine 293 of the human Pyruvate Dehydrogenase E1-alpha subunit protein.

Modification

p Ser293

Localization

Mitochondrion matrix

Specificity

This is specific for the phosphorylated Serine 293 form of the PDHE1 alpha protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

43 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images

Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody - BSA Free [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - MEF were treated with 10 uM BA or DMSO (0.1%) for 16 h before they were subjected to a glycolysis stress test as described under "Materials and Methods". MEF were treated with DMSO (0.1%, D) and 10 uM BA for the indicated periods of time before total cell lysates were subjected to immunoblot analyses for pPDHE1 (Ser273) and total PDHE1 (molecular weight 43 kDa). Representative blots out of three experiments are shown. The graph below depicts compiled densitometric values of pPDHE/PDHE. Representative blots out of three independent experiments are shown. The graph below depicts compiled densitometric values of GLUT1/actin and GLUT3/actin, respectively. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0115683) licensed under a CC-BY license.
Immunocytochemistry/Immunofluorescence: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] antibody (1:250) was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Detection of PDHE1 alpha [S293] in an in vitro autophosphorylation of PDH complex in response to different stimulants: (-) none, H202-hydrogen peroxide, AR-anhydroretinol, Rol-retinol, RA-retinoic acid, PMA, Lipid-PKC lipid activator. Photo courtesy of Dr. Beatrice Hoyos, Memorial Sloan-Kettering Cancer Center.
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Applications

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 31900478)

Immunocytochemistry/ Immunofluorescence

1:50-1:250

Immunoprecipitation

reported in scientific literature (PMID 27450723)

Knockout Validated

reported in scientific literature (PMID 31742248)

Western Blot

1:1000-1:5000
Application Notes
In Western blot, a band is seen ~43 kDa. It is recommended that BSA rather than non-fat milk be used in membrane blocking and antibody dilution.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 5 reviews rated 4.8 using NB110-93479 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS (pH 7.4)

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Pyruvate Dehydrogenase E1-alpha subunit

Pyruvate dehydrogenase (PDH) complex is localized to mitochondrial matrix, wherein it catalyzes the irreversible oxidative decarboxylation of pyruvate to generate acetyl-CoA, NADH, and CO2. The PDH complex contains three primary enzyme components: pyruvate dehydrogenase (E1 or PDHA- heterotetramer of two alpha- and two beta-subunits), dihydrolipoamide transacetylase (E2 or DLAT), and dihydrolipoamide dehydrogenase (E3 or DLD). The activity of PDH is controlled by specific E1 kinase and phospho-E1-phosphatase enzymes, which respectively inactivate and activate PDH complex by phosphorylation and dephosphorylation of E1alpha-subunit's serine residues. PDK family kinases mediated phosphorylation at Ser-293 blocks the access to active site leading to inactivation of the enzyme and vice versa. E3-binding protein (E3BP) is another component which is required for proper interaction of E2 and E3 components, and the overall complex contains 12 copies of E3, 30 copies of E1, 60 copies of E2, and 12 copies of E3BP. Acetyl-CoA generated gets utilized in TCA (citric acid/Krebs cycle) where it reacts with oxaloacetate to form citrate or it may also be used for biosynthesis of fatty acid or cholesterol. Defects in PDHA1 are a cause of pyruvate dehydrogenase E1-alpha deficiency (PDHAD), X-linked Leigh syndrome (X-LS) and it has also been implicated in ageing, glucose intolerance, cancer as well as Alzheimer's disease.

Alternate Names

EC 1.2.4.1, PDHA, PDHE1-A type I, PHE1APDHCE1A, pyruvate dehydrogenase (lipoamide) alpha 1, pyruvate dehydrogenase complex, E1-alpha polypeptide 1, pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial

Gene Symbol

PDHA1

Additional Pyruvate Dehydrogenase E1-alpha subunit Products

Product Documents

Product Datasheets

Certificate of Analysis

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Product Specific Notices

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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